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Nonmigrated cells were removed with cotton swabs, and migrated cells were fixed with ice-cold methanol and stained with 4,6-diamidino-2-phenylindole (DAPI)

Then, TMB chromogen solution (CWBIO) was added, and the absorbance at 450 nm was measured via a SpectraMax plate reader (Molecular Devices)

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Subjects received subcutaneous injections of Melanotan II at a starting dose of 0.01 mg/kg daily (Monday through Friday) for two consecutive weeks